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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 林璧鳳(Bi-Fong Lin) | |
dc.contributor.author | Ya-Chun Yang | en |
dc.contributor.author | 楊雅珺 | zh_TW |
dc.date.accessioned | 2021-06-08T00:06:16Z | - |
dc.date.copyright | 2013-08-26 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-13 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17312 | - |
dc.description.abstract | 本研究擬探討維生素對小鼠B細胞活性及調節功能的影響,以體外 (in vitro) 細胞培養模式,利用anti-mouse CD45R/B220 magnetic particles 自6 ~ 10週齡BALB/c小鼠之初代脾臟、腹腔及皮耶氏體 (Peyer’s patch) 細胞分離B細胞,亦由腹腔細胞培養而得之B-1細胞,分別以維生素A、D、E及葉酸預處理1小時再以LPS刺激48小時,分析細胞存活率、細胞激素及抗體的分泌。接著以qPCR方法分析小鼠脾臟B細胞中IL-10細胞激素之基因表現,亦以細胞內染分析維生素對調節型B細胞百分比的影響。最後以維生素培養的B細胞與經anti-CD3/anti-CD28活化的CD4+ T細胞共培養,探討經維生素培養的B細胞對T細胞增生能力的影響。
結果顯示,維生素D可抑制脾臟、腹腔B細胞及B-1細胞IL-6、IL-10及IgM的分泌,並抑制皮耶氏體B細胞IL-6及TGF-β1分泌的趨勢,且顯著促進脾臟B細胞IL-10 mRNA的表現;缺乏葉酸或補充10倍葉酸之AIN-76飼料餵食5週的BALB/c小鼠,脾臟B細胞在LPS刺激下,缺乏或補充葉酸皆顯著降低IgM的分泌,且IL-6及IL-10的分泌隨飲食中葉酸的增加而增的趨勢。維生素A可增加LPS刺激下脾臟B細胞IL-6 並抑制IL-10及IgM的分泌,且腹腔B細胞以及B-1細胞結果趨勢也相同,並抑制脾臟B細胞IL-10 mRNA的表現;但是,維生素A可增加脾臟B細胞中CD1dhiCD5+細胞比例,即所謂B10細胞;當IL-21存在時,維生素A可增加脾臟B細胞IL-10之分泌量及IL-10+細胞比例;將與IL-21及維生素A培養的B細胞與CD4+ T細胞共培養後,可顯著抑制T細胞之增生。維生素E可增加腹腔B細胞IL-6、IL-10的分泌,但在脾臟B細胞及B-1細胞中趨勢相反,可促進脾臟B細胞IL-10 mRNA的表現;將與維生素E培養45小時之後的B細胞與CD4+ T細胞共培養後,可顯著促進T細胞增生,若B細胞培養時有IL-21存在,則可顯著抑制T細胞增生。 綜合以上結果,維生素可顯著影響小鼠B細胞亞群的活性,其中維生素A與維生素E在IL-21存在時,可增進B細胞的調節功能,並顯著抑制T細胞的增生。 | zh_TW |
dc.description.abstract | In order to investigate the effects of vitamins on mouse B cells activity and regulatory function, we isolated B cells from primary splenic, peritoneal and Peyer’s patch cells of 6 to 10-week-old BALB/c mice by using anti-mouse CD45R/B220 magnetic particles, or B-1 cells derived from primary peritoneal cell cultures. B cells were pretreated with vitamin A, D, E or folic acid for 1 hour following LPS stimulation for 48 hours. The cell viability, cytokine and immunoglobulin levels were analyzed. IL-10 cytokine gene expression of splenic B cells was analyzed by qPCR. We also analyzed the effect of vitamin on percentages of regulatory B cells in splenic B cells by intracellular staining, and on the regulatory function of B cells by pretreating B cells with vitamin and then co-culturing with CD4+ T cells for T cell proliferation using 3H-thymidine incorporation assay.
The results showed that vitamin D inhibits the IL-6, IL-10, and IgM secretions from spleen and peritoneal B cells, as well as B-1 cells, and also inhibits IL-6 and TGF-β1secretion of Peyer’s patch B cells. Folate supplement had little effects on B-cell subsets activity, only the high-dose folic acid increased spontaneous IL-6 and IL-10 secretion of peritoneal B cells. Folate deficiency tended to increase peritoneal B cells and B-1 cells viability, increase TGF-β1 secretion in splenic, peritoneal and Peyer's patch B cells, and increase IL-10 mRNA expression of splenic B cells. Vitamin A increased IL-6, decreased IL-10, IgM secretions of splenic B cells under LPS stimulation, and the same trend was observed in peritoneal B cells and B-1 cells culture. Vitamin A also decreased IL-10 mRNA expression in spleinc B cells, but increased CD1dhiCD5+ cells percentage in splenic B cells. In the presence of IL-21, vitamin A increased IL-10 secretion and IL-10+ cells percentage in splenic B cells. B cells cultured with IL-21 and vitamin A following co-cultured with CD4+ T cells significantly inhibited T cells proliferation. Vitamin E increased IL-6, IL-10 secretion in peritoneal B cells, but in splenic B cells and B-1 cells showed opposite trend. Vitamin E also increased IL-10 mRNA expression in splenic B cells. B cells cultured with high level of vitamin E for 45 hours can promote T cells proliferation after co-cultured with CD4+ T cells. But if B cells cultured with IL-21 and vitamin E following co-cultured with CD4+ T cells, can significantly inhibit CD4+ T cells proliferation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T00:06:16Z (GMT). No. of bitstreams: 1 ntu-102-R00b22026-1.pdf: 5023635 bytes, checksum: 4da6a075643d321c612af2b3523eebd9 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要 I
Abstract III 總目錄 V 圖目錄 X 表目錄 XIII 縮寫對照表 XIV 第一章、緒論 1 第一節、 文獻回顧 1 第二節、 研究動機與大綱 22 一、 研究動機 22 二、 研究大綱 23 第二章、維生素對小鼠不同組織B細胞活性之影響 24 第一節、 前言 24 第二節、 材料與方法 25 一、 細胞實驗設計與流程 25 二、 動物飼養 27 三、 腹腔細胞之取得 27 四、 脾臟細胞之取得 28 五、 皮耶氏體 (Peyer’s patch) 細胞之取得 28 六、 初代B細胞之分離與培養 29 七、 DNA 細胞存活率測定 30 八、 抗體分泌量之分析 30 九、 細胞激素分泌量之分析 31 十、 統計方法 34 第三節、 結果 35 一、 BALB/c 小鼠之各器官組織B細胞所占百分比 35 二、 維生素A對 BALB/c 小鼠B細胞活性之影響 36 三、 維生素D對 BALB/c 小鼠B細胞活性之影響 39 四、 維生素E對 BALB/c 小鼠B細胞活性之影響 42 五、 葉酸對 BALB/c 小鼠B細胞活性之影響 45 六、 缺乏葉酸對 BALB/c 小鼠B細胞活性之影響 48 第四節、 討論 52 一、 不同組織B細胞之活性 52 二、 皮耶氏體B細胞活性探討 52 三、 B細胞分泌 TGF-β1 之能力 53 四、 維生素A對 BALB/c 小鼠B細胞的活性影響 54 五、 維生素D對 BALB/c 小鼠B細胞的活性影響 55 六、 維生素E對 BALB/c 小鼠B細胞的活性影響 56 七、 葉酸對 BALB/c 小鼠B細胞的活性影響 57 第三章、維生素對小鼠 B-1 細胞活性之影響 58 第一節、 前言 58 第二節、 材料與方法 59 一、 細胞實驗設計與流程 59 二、 B-1 細胞之取得 60 三、 B-1 細胞之亞群分佈分析 60 四、 分析項目與統計方法 61 第三節、 結果 62 一、 B-1 細胞亞群分析 62 二、 維生素A對 BALB/c 小鼠 B-1 細胞活性之影響 63 三、 維生素D對 BALB/c 小鼠 B-1 細胞活性之影響 64 四、 維生素E對 BALB/c 小鼠 B-1 細胞活性之影響 65 五、 缺乏葉酸對BALB/c 小鼠 B-1 細胞活性之影響 66 第四節、 討論 67 一、 B-1 細胞之各亞群細胞百分比 67 二、 B-1 細胞之細胞激素以及抗體分泌情況 67 三、 不同維生素對 BALB/c 小鼠 B-1 細胞之影響 67 四、 B-1 細胞取得之困難點 69 第四章、維生素對小鼠脾臟B細胞之細胞激素基因表現的影響 71 第一節、 前言 71 第二節、 材料與方法 72 一、 細胞實驗設計與流程 72 二、 Quantitative PCR 法分析細胞激素基因表現 73 三、 統計方法 75 第三節、 結果 76 一、 維生素A對小鼠脾臟B細胞之細胞激素基因表現的影響 76 二、 維生素D對小鼠脾臟B細胞中細胞激素基因表現之影響 78 三、 維生素E對小鼠脾臟B細胞中細胞激素基因表現之影響 80 四、 缺乏葉酸對小鼠脾臟B細胞中細胞激素基因表現之影響 82 第四節、 討論 84 一、 添加 IL-21 對小鼠B細胞的刺激作用以及鼠種差異 84 二、 維生素對小鼠B細胞 IL-10 mRNA 表現量之影響 84 三、 維生素對小鼠B細胞 IL-24 mRNA 表現量之影響 85 四、 維生素對小鼠B細胞 IL-31 mRNA 表現量之影響 85 五、 維生素對小鼠B細胞 IL-22 mRNA 表現量之影響 86 六、 維生素對小鼠B細胞 IL-27 mRNA 表現量之影響 86 七、 維生素對小鼠B細胞 IL-33 mRNA 表現量之影響 87 八、 總結維生素對小鼠B細胞之細胞激素基因表現影響 87 第五章、維生素對小鼠脾臟B細胞之功能性影響探討 88 第一節、 前言 88 第二節、 材料與方法 89 一、 B10 細胞實驗設計與流程 89 二、 B10 細胞培養之實驗流程 90 三、 免疫細胞表型分析 90 四、 初代 CD4+ T 細胞之分離 92 五、 B細胞與T細胞共培養實驗 93 六、 細胞之增生測定 94 七、 經 FBS 結合處理之維生素E對小鼠脾臟B細胞活性影響 95 第三節、 結果 96 一、 維生素A對小鼠脾臟B細胞中調節型B細胞百分比之影響 96 二、 維生素E對 BALB/c 小鼠脾臟B細胞中 B10 細胞百分比之影響 98 三、 維生素A培養之初代脾臟B細胞對 CD4+ T 細胞增生能力的影響 99 四、 維生素E培養之初代脾臟B細胞對 CD4+ T 細胞增生能力的影響 100 五、 FBS 結合處理維生素E對小鼠脾臟B細胞活性之影響 101 六、 FBS 結合處理維生素E培養之初代脾臟B細胞對 CD4+ T 細胞增生能力的影響 102 第四節、 討論 103 一、 L+PIM 以及 IL-21 對 B10 細胞百分比的影響 103 二、 維生素A對B10細胞的影響 103 三、 維生素E對B10細胞的影響 104 四、 以維生素A培養的B細胞對活化之T細胞增生的影響 104 五、 以維生素E培養的B細胞對活化之T細胞增生的影響 105 六、 比較FBS 結合處理之維生素E對小鼠B細胞活性之影響 106 七、 以經 FBS 結合處理的維生素E培養的B細胞對活化之T細胞增生的影響 106 第六章、綜合討論與總結 107 第一節、 綜合討論 107 一、 維生素A對小鼠B細胞活性之影響 107 二、 維生素D對小鼠B細胞活性之影響 108 三、 以維生素E培養之小鼠B細胞對活化的 CD4+ T 細胞增生情形之影響 109 四、 補充或缺乏葉酸對小鼠B細胞活性之影響 111 五、 B-1 細胞培養模式之建立以及 B-1 細胞於免疫系統中可能扮演之調節角色 111 六、 維生素對小鼠B細胞 IL-10表現與分泌之影響 112 第二節、 總結 114 參考文獻 115 | |
dc.language.iso | zh-TW | |
dc.title | 維生素對小鼠B細胞活性與調節功能之影響 | zh_TW |
dc.title | The effects of vitamins on mouse B cells activity and regulatory functions | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 江伯倫(Bor-Luen Chiang),黃青真,吳文勉,洪永瀚 | |
dc.subject.keyword | 維生素A,維生素E,B-1細胞,調節型B細胞,IL-10,IL-21, | zh_TW |
dc.subject.keyword | vitamin A,vitamin E,B-1 cells,regulatory B cells,IL-10,IL-21, | en |
dc.relation.page | 127 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2013-08-13 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-102-1.pdf 目前未授權公開取用 | 4.91 MB | Adobe PDF |
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